Components of TGFβ-signaling pathway in human subcutaneous AT cells.

<p><i>TGFβ R1</i> (<i>ALK5</i>) (A), <i>activinA R1</i> (<i>ACVR1A/ALK2</i>) (B), <i>SMAD2</i> (C), <i>SMAD3</i> (D), <i>FIBRONECTIN</i> (E) and <i>PAI-1</i> (F) transcript levels were determined by real-time PCR in ScAT mature adipocytes (Adip), endothelial cells (EC), progenitor cells (Prog) and ATM. Values are means ± SEM (AU, arbitrary units) of 5 to 27 independent. * P<0.05, ** P<0.01 and *** P<0.001 between cell types.</p

Hypoxia and mature adipocyte-derived factors affect the remodeling phenotype of human subcutaneous AT macrophages.

<p>Transcript levels of angiogenic and matrix remodeling/fibrotic factors were determined by real-time PCR analyses on ScATM cultured for 24 h (A) in hypoxic conditions (1% O₂, n = 5) and (B) in mature subcutaneous adipocyte-conditioned media (CM, n = 5). Values, expressed as a percentage of the control, are means ± SEM. * P<0.05 and ** P<0.01 <i>vs</i> control conditions.</p

Obesity is associated with increased expression of myofibroblast markers in subcutaneous AT progenitor cells.

<p>A and B, Representative photomicrograph of immunohistochemistry of whole ScAT staining: α-SMA (red), CD34 (green) and nuclei (Hoechst 33242/blue) (n = 9). White scale corresponds to 50 µm. C, <i>SNAIL</i> and <i>SLUG</i> transcript levels were determined by real-time PCR in immunoselected ScAT progenitor cells from 7 non obese (Nob) and 8 obese (Ob) individuals. Values are means ± SEM (AU, arbitrary units). * P<0.05 and ** P<0.01 <i>vs</i> Nob.</p

TGFβ1 and subcutaneous AT macrophage-conditioned media induce a myofibroblast-like phenotype in human AT progenitor cells.

<p>A and B, Transcript levels of <i>SNAIL</i>, <i>SLUG</i> and <i>INHBA</i>/activinA were determined by real-time PCR analyses in ScAT progenitor cells treated for 24 h with basal medium (i.e, control, n = 4 to 5), with TGFβ1 (5 ng/ml, n = 4) or with ScATM-CM (n = 6). Results are expressed as percentage of the control and are means ± SEM. * P<0.05 and ** P<0.01 <i>vs</i> control media. C, Representative photomicrograph of immunocytochemical staining for α-SMA (magnification ×10) of ScAT progenitors cells (n = 3) that were cultured for 48 h with ScATM-CM (n = 5) in the presence or not of neutralizing antibodies directed against TGFβ (1 µg/ml), activinA (1 µg/ml) or both (A+T, 1 µg/ml each). White scale corresponds to 50 µm. D, Number of α-SMA⁺ foci per 100 nuclei. Values are expressed as a percentage of ScATM-CM and are means ± SEM. * P<0.05 <i>vs</i> ScATM-CM, n = 3. E, Transcript levels of <i>SNAIL</i>, <i>SLUG</i> and <i>α-SMA</i> were determined by real-time PCR analyses in hMADS treated or not with activinA (100 ng/ml) for 24 h (n = 4). Values are expressed as fold increase of the controland are means ± SEM. * P<0.05 and *** P<0.001 <i>vs</i> control media.</p

AT location affects the remodeling phenotype of human AT macrophages in obese individuals.

<p>A, Transcript levels of angiogenic and matrix remodeling/fibrotic factors were determined by real-time PCR analyses of ATM immunoselected from paired biopsies of subcutaneous (Sc) and omental AT (Om). Results are expressed as fold differences between Om and Sc and are means ± SEM (n = 22 subjects, mean BMI 43.9±1.4 kg/m²). Open bars: genes up-regulated, and solid bars: genes down-regulated, in OmATM <i>vs</i> ScATM. B, Transcript levels of <i>HIF-1α</i> and <i>-2α</i> determined by real-time PCR analyses of ScATM and OmATM. Values are means ± SEM (AU, arbitrary unit) of the 22 paired biopsies. * P<0.05 and *** P<0.001 <i>vs</i> Om.</p

TGFβ1 and subcutaneous AT macrophage-conditioned media induce α-SMA expression in human AT progenitor cells.

<p>Representative photomicrograph of immunocytochemical staining for α-SMA (red) of ScAT progenitors cells (n = 4) cultured for 48 h with basal medium (i.e, control), with TGFβ1 (5 ng/ml, n = 4, upper panel) or with ScATM-CM (n = 6, middle and lower panels). Nuclei were stained with Hoechst 33242 (blue). Magnification ×20 and ×40. White scale corresponds to 50 µm.</p

Obesity status affects the remodeling phenotype of human subcutaneous AT macrophages.

<p>ATM were immunoselected from subcutaneous AT from 18 non obese individuals (Nob, mean BMI 25.4±0.6 kg/m²) and 22 obese individuals (Ob, mean BMI 43.9±1.4 kg/m²) and the transcript levels of angiogenic and matrix remodeling/fibrotic factors were determined by real-time PCR analyses. Values are means ± SEM (AU, arbitrary unit). * P<0.05, *** P<0.001 <i>vs</i> Nob. IL, interleukin; MCP-1, monocyte chemotactic protein 1; MMP, matrix metalloproteinase; VEGF, vascular endothelial growth factor; LYVE-1, lymphatic vessel endothelial hyaluronan receptor 1.</p